Extraction of DNA from Malaria-Infected Erythrocytes Using Isotachophoresis

被引:37
作者
Marshall, Lewis A. [2 ]
Han, Crystal M. [1 ]
Santiago, Juan G. [1 ]
机构
[1] Stanford Univ, Dept Mech Engn, Stanford, CA 94305 USA
[2] Stanford Univ, Dept Chem Engn, Stanford, CA 94305 USA
关键词
PURIFICATION; PCR;
D O I
10.1021/ac202567j
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
We demonstrate a technique for purification of nucleic acids from malaria parasites infecting human erythrocytes using isotachophoresis (ITP). We release nucleic acids from malaria-infected erythrocytes by lysing with heat and proteinase K for 10 min and immediately, thereafter, load sample onto a capillary device. We study the effect of temperature on lysis efficiency. We also implement pressure-driven counterflow during ITP extraction to extend focusing time and increase nucleic acid yield. We show that the purified genomic DNA samples are compatible with polymerase chain reaction (PCR) and demonstrate a clinically relevant limit of detection of 0.5 parasites per nanoliter using quantitative PCR.
引用
收藏
页码:9715 / 9718
页数:4
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